Single-molecule microfluidic assay for prostate-specific antigen based on magnetic beads and upconversion nanoparticles

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Authors

SKLENÁROVÁ Dorota HLAVÁČEK Antonín KŘIVÁNKOVÁ Jana BRANDMEIER Julian WEISOVÁ Julie ŘIHÁČEK Michal GORRIS Hans-Heiner SKLÁDAL Petr FARKA Zdeněk

Year of publication 2024
Type Article in Periodical
Magazine / Source Lab on a Chip
MU Faculty or unit

Faculty of Science

Citation
Web https://pubs.rsc.org/en/content/articlelanding/2024/lc/d4lc00346b
Doi http://dx.doi.org/10.1039/D4LC00346B
Keywords Photon-upconversion nanoparticle; Immunoassay; Microfluidics; Prostate-specific antigen
Description Early-stage diagnosis of prostatic carcinoma is essential for successful treatment and, thus, significant prognosis improvement. In laboratory practice, the standard non-invasive diagnostic approach is the immunochemical detection of the associated biomarker, prostate-specific antigen (PSA). Ultrasensitive detection of PSA is essential for both diagnostic and recurrence monitoring purposes. To achieve exceptional sensitivity, we have developed a microfluidic device with a flow-through cell for single-molecule analysis using photon-upconversion nanoparticles (UCNPs) as a detection label. For this purpose, magnetic microparticles (MBs) were first optimized for the capture and preconcentration of PSA and then used to implement a bead-based upconversion-linked immunoassay (ULISA) in the microfluidic device. The digital readout based on counting single nanoparticle-labeled PSA molecules on MBs enabled a detection limit of 1.04 pg mL-1 (36 fM) in 50% fetal bovine serum, which is an 11-fold improvement over the respective analog MB-based ULISA. The microfluidic technique conferred several other advantages, such as easy implementation and the potential for achieving high-throughput analysis. Finally, it was proven that the microfluidic setup is suitable for clinical sample analysis, showing a good correlation with a reference electrochemiluminescence assay (recovery rates between 97% and 105%).
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